Download

1- Making Density Gradients






Advertisement
/ 24 []
Download Presentation
Comments
montana
From:
|  
(1313) |   (0) |   (0)
Views: 346 | Added:
Rate Presentation: 1 0
Description:
1- Making Density Gradients. Pre-formed discontinuous gradients Pre-formed continuous gradients Self-generated gradients. 2- Density Gradient Harvesting. C. A. B. Discontinuous gradient by overlayering. A. B. C. D. Discontinuous gradient by underlayering. density. top. bottom.
1- Making Density Gradients

An Image/Link below is provided (as is) to

Download Policy: Content on the Website is provided to you AS IS for your information and personal use only and may not be sold or licensed nor shared on other sites. SlideServe reserves the right to change this policy at anytime. While downloading, If for some reason you are not able to download a presentation, the publisher may have deleted the file from their server.











- - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - E N D - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -




1 making density gradients l.jpgSlide 1

1- Making Density Gradients

  • Pre-formed discontinuous gradients

  • Pre-formed continuous gradients

  • Self-generated gradients

2- Density Gradient Harvesting

Discontinuous gradient by overlayering l.jpgSlide 2

C

A

B

Discontinuous gradient by overlayering

Discontinuous gradient by underlayering l.jpgSlide 3

A

B

C

D

Discontinuous gradient by underlayering

Diffusion of a discontinuous gradient l.jpgSlide 4

density

top

bottom

Diffusion of a discontinuous gradient

Rapid formation of continuous from discontinuous gradient l.jpgSlide 5

Discontinuous

Continuous

45-60 min

Rapid formation of continuous from discontinuous gradient

Two chamber device for continuous gradients l.jpgSlide 6

A

B

T

SB

SB

P

M

Two-chamber device for continuous gradients

The gradient master l.jpgSlide 7

B

The Gradient Master

Gradient master profiles from 10 and 40 iodixanol at 80 and 20 rpm effect of time l.jpgSlide 8

Density (g/ml)

2 min

1.13

2.5 min

3.0 min

1.11

1.09

1.07

1

3

5

7

9

Fraction Number

Gradient Master profiles from 10% and 40% iodixanol at 80° and 20 rpm: effect of time

Swinging bucket rotor l.jpgSlide 9

axis of rotation

g

At rest

Spinning

g

Swinging-bucket rotor

Fixed angle and vertical rotors l.jpgSlide 10

Fixed-angle

Vertical

g

1g

Fixed-angle and vertical rotors

Sedimentation path length of rotors l.jpgSlide 11

Swinging-bucket

Axis of

rotation

Vertical

Sedimentation path length of rotors

Sample gradient in vertical rotor l.jpgSlide 12

A

B

Axis of rotation

Sample/gradient in vertical rotor

Self generated gradients l.jpgSlide 13

time

g

Self-generated gradients

  • At 360,000gav iodixanol molecules will start to sediment

Self generated gradient strategy l.jpgSlide 14

350,000g

350,000g

1g

1g

Self-generated gradient strategy

Near vertical rotors are best of all l.jpgSlide 15

Near-vertical rotors are best of all

Iodixanol self generated gradient requirements l.jpgSlide 16

Iodixanol self-generated gradient requirements

  • Rotors with a short sedimentation path length ~ 17 mm

  • RCF of 180,000-350,000gav

Effect of time on iodixanol self generated gradient in the beckman tln100 at 365 000g l.jpgSlide 17

Density (g/ml)

15% iodixanol

1.30

1 h

3 h

1.25

20% iodixanol

1.20

1 h

3 h

1.15

1.10

1.05

1.00

1

3

5

7

9

11

13

15

Fraction Number

Effect of time on iodixanol self-generated gradient in the Beckman TLN100 at 365,000g

Effect of rcf and iodixanol in beckman vti65 1 3 h l.jpgSlide 18

1.3

15% (170,000g)

30% (170,000g)

15% (353,000g)

1.2

30% (353,000g)

Density (g/ml)

1.1

1.0

1

3

5

7

9

11

Fraction Number

Effect of RCF and [iodixanol] in Beckman VTi65.1 (3 h)

Removal of banded material using a syringe l.jpgSlide 19

Removal of banded material using a syringe

Collecting a band from a sealed tube l.jpgSlide 20

Collecting a band from a sealed tube

Harvest gradient into equal volume fractions by tube puncture l.jpgSlide 21

Gilson Fraction

Collector

P

96-well plate

Harvest gradient into equal volume fractions by tube puncture

Upward displacement l.jpgSlide 22

Perfluorodecalin

Perfluorodecalin

P

P

Upward displacement

Labconco auto densi flow collects from meniscus l.jpgSlide 23

P

P

P

P

M

B

EP

C

1

2

3

4

Labconco Auto Densi-flow collects from meniscus

Labconco auto densi flow linked to a gilson fc205 fraction collector l.jpgSlide 24

MB

Labconco Auto Densi-flow linked to a Gilson FC205 Fraction Collector


Copyright © 2014 SlideServe. All rights reserved | Powered By DigitalOfficePro